The prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) is an excellent model to study the molecular and cellular biology of arenaviruses that cause important and severe human diseases like Lassa fever virus (LFV) and the South American viral hemorrhagic fevers (SAHF). In addition, weaponized forms of these viruses pose a real threat as agents of bioterrorism. No licensed vaccine is available in the US, and currently there is not efficacious therapy to treat these viral infections. Therefore the importance of developing novel effective antiviral drugs to combat pathogenic arenaviruses. We have developed a reverse genetic system for LCMV. This system provides us with a robust platform for the development of novel strategies to target specific steps of the Arenavirus life cycle. We have molecularly characterized the Arenavirus genome promoter. Disruption of the interaction between the viral polymerase and promoter is predicted to abrogate virus multiplication. RNA molecules can form intricate structures that can be targeted by selected specific high-affinity antagonists. This proposal will explore the use of aminoglycoside-based small molecules to target and functionally disrupt the Arenavirus genome promoter, thus inhibiting virus infection. Aim 1 is the screening of combinatorial aminoglycoside libraries to identify small molecules that bind to the LCMV genome promoter. For this, biotin-RNA conjugates of the LCMV promoter will be attached to streptavidin-coated sensor chips and used as a ligand to screen the libraries using surface plasmon resonance. Aim 2 will assess the antiviral activity of binder molecules (Aim 1). Selected aminoglycosides will be tested for: (i) their effects on RNA synthesis mediated by the LCMV polymerase using our LCMV minigenome system; (ii) their ability to inhibit LCMV multiplication in cultured cells. Finlly, Aim 3 will assess the generation of viral variants resistant to antiviral aminoglycosides. Emergence of resistant variants will be assessed based on production of infectious virus and intracellular levels of virus RNA synthesis during serial passages in the presence of aminoglycosides with anti-LCMV activity. [unreadable] [unreadable]